LOCUS       40924_13560        8114 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCS2+MT-gli2act, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8114)
  AUTHORS   Cerrizuela S, Vega-Lopez GA, Palacio MB, Aybar MJ.
  TITLE     Gli2 is required for the induction and migration of Xenopus laevis 
            neural crest
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8114)
  AUTHORS   Cerrizuela S, Vega-Lopez GA, Palacio MB, Aybar MJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-JUL-2017) Developmental Biology, Instituto Superior de
            Investigaciones Biologicas (INSIBIO, CONICET), Chacabuco 461, San 
            Miguel de Tucuman, Tucuman T4000ILI, Argentina
REFERENCE   3  (bases 1 to 8114)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8114)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (04-JUL-2017) Developmental Biology, Instituto Superior de 
            Investigaciones Biologicas (INSIBIO, CONICET), Chacabuco 461, San 
            Miguel de Tucuman, Tucuman T4000ILI, Argentina"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8114
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             6..4067
                     /codon_start=1
                     /gene="Gli2"
                     /product="Gli2act"
                     /label=Gli2
                     /note="activator form of Gli2 protein"
                     /protein_id="ATQ62666.1"
                     /translation="MGGGALSISPLSDASIDLQTMIRTSPNSLVAYINNSRSSSAASGS
                     YGHLSAGAISPAFSFPIHKPCSLSAALSQQRSLSSSFGHTPLLHPSPTFASRQQGALTS
                     ANPAPPSNNSSAPDSVLNKVSSESAVSSTVNQVIHKRSKVKTEEEADSVRFPQPPDHLT
                     DLKEDLDKDECKQEPEHIYETNCHWDGCSKEFDTQDQLVHHINNDHIHGEKKEFVCRWQ
                     DCSREQKPFKAQYMLVVHMRRHTGEKPHKCTFEGCFKAYSRLENLKTHLRSHTGEKPYV
                     CDHEGCNKAFSNASDRAKHQNRTHSNEKPYICKVPGCTKRYTDPSSLRKHVKTVHGPEA
                     HVTKKHRNDIIQKPSLPKENGDNEASAKLSGREHSDSVSRDQEHCLQTRTIKTEDNMMH
                     QSSPGGQSSCSSEPSPYGNTNNIDSGVDVSLAWQGSLGDLFGLEETSPVVDSTVSSWQR
                     SGRPATPETQRIHSAETGTAEEREIKDNERFLLIYEPNATCQNTRLPTISANGFDVIGV
                     PSSVLINPRAIELSMNDVTMMNQLNERRDSTSSTLSSAYTSRRSSGISPYFSSRRSSET
                     SQFGGRLNNSSSADSYDPISTDASRRSSEASQHSGLPNLLNLTPAQHYRLKAKYAAATG
                     GPPPTPLPNMDRIGLRNKLSLMDGADFPLPPFRQLPVPRRCSDGGGNAGLTPMYPHEIP
                     GNNSRRASDPVRRTAGIDDKPLPRFSRFHSMNSMNTLHPPSLSERRNGGLQHYTCSDGG
                     LHRHVYSPRPPSISENVAMEAISCDADVPGGDDDLMLPDDVVQYIRSQNREAPEQNLQT
                     EYSSPARNLQSNTKSFHNNTPEQPRAPGAYLSRNFPALAECLGQTANMQDNNMPVQWNE
                     VSSGTVDVSDLPKQQFAAGNLAVVQQKQNFAQYQSFNQAPMQRAHNIMGQGQESVQRNI
                     SVNGQRFNYLQQRQQQMSQCQIVSSDFIPQQRYSQSQSMLSSRAMQEGQSQISPSCNNM
                     VERPGVHTHAAPSNTLNHQRLAVHGAPTQGFANNFSVNQDGLHPPNAYTVQPQKNGLEP
                     QQNTLGMSGQAFNHGMIQPRPPAAPHPPNRPRNIHPVHHPPYMRSPHPVSELSPGQQTA
                     EATPKRTSENTDPTPKDNNLLYYSGQIHMYEPNGNFGSGIDCTVRQLPTMPSPGANQVT
                     STVDSQGLEHPPVIDFDAMMDDGDHSSLMSGTLSPGLLQSFSQTSSRLTTPRNSLTLPS
                     IPAGINNMAIGDMSSMLTTLAEESKFLNLNRFKAMEQKLISEEDLNEMEQKLISEEDLN
                     EMEQKLISEEDLNEMEQKLISEEDLNEMEQKLISEEDLNEMESLGDLTMEQKLISEEDL
                     NSRPLEPLEL"
     gene            6..4067
                     /gene="Gli2"
                     /label=Gli2
     sig_peptide     3783..4037
                     /gene="Gli2"
     CDS             3786..3815
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3825..3854
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3864..3893
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3903..3932
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3942..3971
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4005..4034
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(4062..4079)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      complement(4063..4079)
                     /regulatory_class="promoter"
     polyA_signal    complement(4084..4218)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(4331..4349)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4370..4386)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4394..4410)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4418..4448)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4463..4484)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4772..5360)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5534..6391)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6392..6496)
                     /label=AmpR promoter
     rep_origin      complement(6522..6977)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        7051..8035
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus
                     major immediate early transcription unit IE94"
     promoter        8070..8088
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 gatccatggg gggaggggcc ctttccatct ctcctctgtc cgatgccagc attgacctgc
       61 agaccatgat ccgcacctcc cccaactcgc tggttgctta catcaacaac tctcggagca
      121 gctcagccgc cagcgggtcc tacgggcacc tttctgcagg agcaataagc ccggcgttta
      181 gcttccccat ccataaaccc tgtagcctat cagcagctct gagccagcag cgtagcctga
      241 gttcctcatt tggacacacc cctttgctcc acccatcgcc gacgtttgca tcccgccagc
      301 agggagcact cacatcagcc aatccagctc ctcccagcaa caacagctcc gcccctgact
      361 ccgtcctgaa caaggtgagc agtgagtctg cggtgagcag tacagtcaat caggtgatac
      421 acaaacgcag caaagtgaag actgaagaag aggccgattc agtgcgattc ccccagcctc
      481 cggaccacct gactgatctg aaagaggatt tggataagga tgagtgcaag caggagcctg
      541 aacatatata cgagaccaac tgccactggg acggctgcag caaggagttt gatacccagg
      601 atcagctcgt acatcacatc aataatgacc atatccatgg ggagaagaag gagtttgtgt
      661 gtcgctggca ggactgctcc cgggagcaga aaccattcaa ggcgcagtat atgctggtgg
      721 ttcacatgag aaggcacacg ggggagaaac cacataaatg cacctttgaa ggttgtttca
      781 aggcctattc caggctggag aacctgaaga cgcacttgcg ttcacacacc ggggagaagc
      841 catatgtctg tgaccacgag ggatgcaata aggcgttttc caatgcgtct gatcgagcca
      901 aacatcagaa cagaacccat tccaatgaga aaccgtacat atgtaaggtc ccaggctgca
      961 caaaacgtta cacggacccc agctctctca ggaaacacgt taaaactgta catggacctg
     1021 aggcccatgt gaccaagaag caccgcaatg acatcatcca aaaaccttca ctacccaagg
     1081 agaacgggga caatgaagct agtgccaaac tcagcggtag ggagcactcc gacagtgtct
     1141 ctagagacca ggagcactgc ttgcagacca gaaccatcaa aacagaagat aacatgatgc
     1201 atcagtccag ccccggtggg cagtcgtcat gcagcagtga gccttcccca tatggcaaca
     1261 ccaacaacat tgacagtgga gtagatgtga gccttgcatg gcaggggagc ttgggagacc
     1321 tgtttggctt ggaggagacc agcccggtgg tagactccac agtatcatcc tggcagcgca
     1381 gtggtcggcc tgcaactccg gaaacacagc gtatccacag tgcagagact ggaacagctg
     1441 aagaaagaga aattaaagac aatgaaagat tcctgctcat ttatgaaccc aatgccacct
     1501 gccagaacac ccggctcccg accatcagtg caaatggttt cgatgtaata ggtgtcccca
     1561 gcagtgtgct aatcaacccc agagcaatag aactgtccat gaatgatgtg accatgatga
     1621 accaattgaa tgagcgccgt gacagcacca gcagcaccct cagctcagca tacaccagcc
     1681 gcaggtcctc cggcatctct ccttacttct ccagtcgccg ctcaagtgaa acctcacagt
     1741 ttggaggccg cttgaataat tccagctctg cagattctta cgatcccatt tccaccgatg
     1801 cctctaggcg ctctagtgag gccagccagc acagtggact tcctaatctt ctcaacctca
     1861 ctcctgctca gcattacaga ttaaaggcca agtatgctgc agcaactggt ggtccaccac
     1921 ccactcctct tcccaacatg gataggatag gtttgagaaa caagctgtcc ctgatggatg
     1981 gagcagactt tcctttgcct cccttccggc aacttcctgt tccaagacgt tgtagtgatg
     2041 gtggtggaaa tgcaggacta acccccatgt atccccatga aattccgggg aataattcaa
     2101 ggcgggctag tgacccagtc agaagaactg caggtattga tgacaagcca ctcccaaggt
     2161 tcagccgctt tcatagcatg aatagtatga atacattgca ccctccatcc ctatcggaaa
     2221 ggagaaatgg aggcctgcag cactacacct gctctgatgg aggccttcac aggcacgtct
     2281 attctccccg accacccagc attagtgaaa atgttgctat ggaggccata tcttgtgatg
     2341 cagatgtccc tggtggagat gatgacctca tgctcccaga tgatgttgtt cagtacatca
     2401 gatctcagaa cagagaagcc ccagagcaaa acctacagac ggagtacagc agtccagccc
     2461 ggaatctaca aagcaacaca aaatctttcc ataacaatac accagaacaa cccagggccc
     2521 ctggggctta cttgagcagg aactttcctg ctttggctga atgccttggc cagactgcta
     2581 acatgcagga taacaacatg cccgttcagt ggaacgaagt cagttcgggt accgttgatg
     2641 tcagtgactt acccaagcag cagtttgcag caggcaactt agcagttgtt cagcaaaaac
     2701 agaactttgc ccagtatcag agcttcaacc aagcacctat gcagagggct cacaatatta
     2761 tgggtcaggg gcaagagtct gtacagagga atatcagtgt taatggacag cggtttaatt
     2821 atttgcaaca aagacaacag cagatgagcc agtgccaaat tgtcagttct gacttcatcc
     2881 cccaacagag gtacagccag agccagtcaa tgctgagttc aagagcaatg caagaaggcc
     2941 agagccagat ttctccatcc tgtaataaca tggtagagag gcctggggtt cacacacatg
     3001 ctgctccaag caacacactg aatcatcagc gccttgctgt gcacggagca cccacgcagg
     3061 gttttgcaaa taacttctcc gttaaccaag acgggctcca cccacccaac gcctacacag
     3121 tccagcctca gaaaaatgga ttggagccac aacagaacac tcttggaatg tctgggcaag
     3181 cgttcaacca tggaatgatt cagccaagac ccccagctgc cccacacccc cctaacagac
     3241 ccaggaatat acacccggta catcaccctc cttacatgag gagtccgcac cctgtgagtg
     3301 aactgagccc aggccaacaa acagcagagg caacaccaaa acgaaccagt gaaaacactg
     3361 atccaactcc taaagacaat aatttgcttt attattctgg acaaattcac atgtacgagc
     3421 caaatggaaa cttcggttct ggcatagact gcacagtacg gcaactaccc accatgccgt
     3481 ctcctggggc aaatcaggtc acaagcactg tagactccca aggtcttgag cacccccctg
     3541 ttatagattt tgacgccatg atggatgatg gggatcactc gagccttatg tcaggaacct
     3601 tgagccctgg cctcctgcag agcttctccc aaacatcctc ccgccttaca actcccagga
     3661 attccctgac gctcccttcc atccctgctg ggataaacaa catggcaata ggagacatga
     3721 gctccatgct cacaacattg gcggaggaaa gcaagtttct caatctgaat cgatttaaag
     3781 ctatggagca aaagctcatt tctgaagagg acttgaatga aatggagcaa aagctcattt
     3841 ctgaagagga cttgaatgaa atggagcaaa agctcatttc tgaagaggac ttgaatgaaa
     3901 tggagcaaaa gctcatttct gaagaggact tgaatgaaat ggagcaaaag ctcatttctg
     3961 aagaggactt gaatgaaatg gagagcttgg gcgacctcac catggagcaa aagctcattt
     4021 ctgaagagga cttgaattca aggcctctcg agcctctaga actatagtga gtcgtattac
     4081 gtagatccag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     4141 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     4201 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     4261 gaggtgtggg aggtttttta attcgcggcc gcggcgccaa tgcattgggc ccggtaccca
     4321 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
     4381 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4441 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4501 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4561 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4621 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4681 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4741 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4801 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4861 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4921 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4981 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5041 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5101 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5161 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     5221 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5281 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5341 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5401 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5461 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5521 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5581 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5641 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5701 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5761 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5821 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5881 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5941 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6001 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6061 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6121 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6181 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6241 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6301 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6361 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6421 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6481 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
     6541 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     6601 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     6661 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     6721 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     6781 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     6841 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     6901 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     6961 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc
     7021 ggtgcgggcc tcttcgctat tacgccagtc gaccatagcc aattcaatat ggcgtatatg
     7081 gactcatgcc aattcaatat ggtggatctg gacctgtgcc aattcaatat ggcgtatatg
     7141 gactcgtgcc aattcaatat ggtggatctg gaccccagcc aattcaatat ggcggacttg
     7201 gcaccatgcc aattcaatat ggcggacttg gcactgtgcc aactggggag gggtctactt
     7261 ggcacggtgc caagtttgag gaggggtctt ggccctgtgc caagtccgcc atattgaatt
     7321 ggcatggtgc caataatggc ggccatattg gctatatgcc aggatcaata tataggcaat
     7381 atccaatatg gccctatgcc aatatggcta ttggccaggt tcaatactat gtattggccc
     7441 tatgccatat agtattccat atatgggttt tcctattgac gtagatagcc cctcccaatg
     7501 ggcggtccca tataccatat atggggcttc ctaataccgc ccatagccac tcccccattg
     7561 acgtcaatgg tctctatata tggtctttcc tattgacgtc atatgggcgg tcctattgac
     7621 gtatatggcg cctcccccat tgacgtcaat tacggtaaat ggcccgcctg gctcaatgcc
     7681 cattgacgtc aataggacca cccaccattg acgtcaatgg gatggctcat tgcccattca
     7741 tatccgttct cacgccccct attgacgtca atgacggtaa atggcccact tggcagtaca
     7801 tcaatatcta ttaatagtaa cttggcaagt acattactat tggaaggacg ccagggtaca
     7861 ttggcagtac tcccattgac gtcaatggcg gtaaatggcc cgcgatggct gccaagtaca
     7921 tccccattga cgtcaatggg gaggggcaat gacgcaaatg ggcgttccat tgacgtaaat
     7981 gggcggtagg cgtgcctaat gggaggtcta tataagcaat gctcgtttag ggaaccgcca
     8041 ttctgcctgg ggacgtcgga gcaagcttga tttaggtgac actatagaat acaagctact
     8101 tgttcttttt gcag
//
